Anti-inflammatory compositions for treating brain inflammation

ABSTRACT

Compositions with synergistic anti-inflammatory effects in inflammatory diseases resulting from activation and consequent degranulation of mast cells and followed by secretion of inflammatory biochemicals from the activated mast cells, the compositions containing one or more of a flavone or flavonoid glycoside a heavily sulfated, non-bovine proteoglycan, an unrefined olive kernel extract that increases absorption of these compositions in various routes of administration, a hexosamine sulfate such as D-glucosamine sulfate, S-adenosylmethionine, a histamine-1 receptor antagonist, a histamine-3 receptor agonist, an antagonist of the actions of CRR, a long-chain unsaturated fatty acid, a phospholipid, Krill oil, a polyamine, glutiramer acetate and interferon. Certain of the present compositions are useful in protecting against the neuropathological components of multiple sclerosis and similar inflammatory neurological diseases.

This application is a continuation of U.S. application Ser. No.11/214,831, filed Aug. 31, 2005, which is a continuation-in-partapplication of U.S. patent application Ser. No. 10/811,826, filed Mar.30, 2004, which is a continuation-in-part application of U.S. patentapplication Ser. No. 09/771,669, filed Jan. 30, 2001, now U.S. Pat. No.6,984,667, which is a continuation-in-part application of U.S. patentapplication Ser. No. 09/056,707, filed Apr. 8, 1998, now U.S. Pat. No.6,689,748.

BACKGROUND OF THE INVENTION

The invention is generally related to the treatment of inflammatoryconditions. More specifically, the invention is related to compositionscontaining inhibitors of mast cell activation and secretion such as aproteoglycan and a flavonoid compound that are designed to be used asdietary supplements or adjuvants to conventional approved medicationsfor the relief of inflammatory conditions, e.g., in the brain as inmultiple sclerosis.

There have been a number of mostly anecdotal reports that theproteoglycan chondroitin sulfate, as well as glucosamine sulfate, aproduct of the intestinal breakdown of proteoglycans, may be helpful inrelieving the pain of osteoarthritis:—Shute N. Aching for an arthritiscure. US News and World Report, Feb. 10, 1997.—Cowley, G. The arthritiscure? Newsweek, Feb. 17, 1997; Foreman J., People, and their pets, toutarthritis remedy. The Boston Globe, Apr. 7, 1997; Tye L. Treatment gainsscientific attention. The Boston Globe, Sep. 25, 2000.

A meta-analysis showed potential therapeutic benefit of chondroitinsulfate and/or glucosamine in osteoarthritis [McAlindon, et al. J. Am.Med. Assn. 283:1469 (2000)], while a double-blind clinical trial withglucosamine showed definite benefits in osteoarthritis with respect toboth pain and radiographic joint appearance [Reginster, et al., Lancet337:252 (2001)]. However, less than 5% of the chondroitin sulfate incommercially available preparations is absorbed orally, because the sizeof the molecule and the degree of sulfation impede its absorption fromthe gastrointestinal tract. Furthermore, such commercial preparationsuse chondroitin sulfate obtained from cow trachea, with the possibledanger of contracting spongioform encephalopathy or “mad cow disease”.In fact, the European Union has banned even cosmetics that containbovine-derived products.

Theoharides, et al., British Journal of Pharmacology 131:1039 (2000)indicated for the first time how proteoglycans, such as chondroitinsulfate, may work. This paper reported that chondroitin sulfate and, toa lesser degree, glucosamine sulfate, inhibit activation of mast cellsthat are known to trigger allergy and asthma. This discovery is thebasis for Theoharides, T. C., U.S. Pat. No. 6,689,748 and Ser. No.09/773,576, filed Feb. 2, 2001.

Mast cells are also now recognized as important causative intermediariesin many painful inflammatory conditions [Galli, N. Eng. J. Med. 328:257(1993); Theoharides, Int. J. Tissue Reactions 18:1 (1996)], such asinterstitial cystitis and irritable bowel syndrome [Theoharides, T. C.,Ann. NY Acad. Sci. 840:619 (1998)], as well as in migraines[Theoharides, T. C., Brain Res. Rev. 49:65 (2005) and possibly multiplesclerosis [Theoharides, T. C., Persp Biol Med. 26:672 (1983),Theoharides, Life Sci. 46:607 (1996), and J. Neuroimmunol. 146:1 (2004).

Mast cells are increasingly implicated in conditions involving inflamedjoints, such as in osteoarthritis and rheumatoid arthritis, throughactivation of local mast cells by, for example, neuropeptides, such asSubstance P. Additional indirect evidence also supports the involvementof mast cells in bone resorption: (a) systemic mastocytosis isinvariably associated with osteoporosis; (b) inhibition of mast cellmediator release reversed lytic bone changes; (c) depletion of mastcells inhibited bone resorption in organ culture; (d) human synovialmast cells were shown to secrete in response to allergic andnon-immunologic stimuli; (e) human mast cells release the cytokine IL-6;and (f) IL-6 has been definitively linked to bone resorption andosteoporosis.

It was shown that chondroitin sulfate's ability to inhibit theactivation of mast cells compliments the inhibitory effects on mast cellactivation of another class of naturally occurring compounds, theflavonoids [Middleton, et al., Pharm. Rev. 52:1 (2000)]. Certain plantflavones (in citrus fruit pulp, seeds, sea weed) are now recognized asanti-allergic, anti-inflammatory, anti-oxidant and cytoprotective withpossible anti-cancer properties. Only some flavonoids, especially thosebelonging to the subclass of flavonols, e.g., quercetin, inhibit mastcell activation.

Quercetin inhibits secretion from human activated mast cells [Kimata, etal., Allergy 30:501 (2000)], and has also been used effectively for thetreatment of chronic prostatitis [Shoskes, et al., Urology 54:960(1999)]. However, other flavonoids may have opposite effects. Use of theterm “bioflavonoids” or “citrus flavonoids” in certain commercialproducts, therefore, provides little information, and may includemolecules that have detrimental effects; for example, soy containsisoflavones that have estrogen-like activity that worsens inflammatoryconditions.

U.S. Pat. No. 6,689,748, and divisional application Ser. No. 09/773,576claim the oral use of proteoglycans, without and with flavonoids, forthe treatment of mast cell activation-induced diseases. Absorption ofthese compositions from the gastrointestinal tract and synergism withother treatment modalities were not addressed in these applications.

Applicant has described the use of antagonists of the action ofCorticotropin Releasing Hormone (“CRH”) (also known as CorticotropinReleasing Factor) in inhibiting myocardial mast cell activation inmyocardial ischemia, in treating stress-induced skin disease (U.S. Pat.No. 6,020,305) and stress-induced migraine headaches (U.S. Pat. No.5,855,884), the contents of which are incorporated herein by reference.The synergistic effects of the compositions of the present inventionthat include antagonists of the actions of CRH on mast cells were notrecognized at the time of the previous studies. The word “antagonists”in connection with CRH is intended herein to include any molecule thatprevents the actions of CRH on target cells, and includes, but is notlimited to, anti-CRH neutralizing antibodies or binding proteins, ormolecules preventing the release of CRH at local sites (see below fordetails).

Applicant has also described a method for treating patients with mastcell derived molecules-induced interstitial cystitis with certainhistamine-I receptor antagonists (Theoharides, U.S. Pat. No. 5,994,357).Treatment of mast cell molecules-induced migraines with histamine-3receptor agonists is the subject of Theoharides U.S. Pat. No. 5,855,884.Histamine-3 receptor agonists as pharmaceutical agents in mastcell-involved diseases are described in Theoharides, U.S. Pat. No.5,831,259. The contents of these three patents are incorporated hereinby reference. At the time of this invention the synergistic effects ofthe present compositions with such antagonists had not yet beenrecognized.

An important need therefore exists for compositions for administrationto human patients being treated for mast cell-induced inflammatorydiseases by various modalities, that are synergistic in that they havestronger effects than the sum of the effects of the individualcomponents, and also synergistic with conventional clinical treatmentsof inflammatory conditions. “Synergistic” is also intended to mean:“coordinated or correlated action by two or more structures ordrugs”[Stedman's Medical Dictionary, 23^(rd) ed., Williams & Wilkins,Baltimore, 1976]. An important need also exists for formulations thatincrease the absorption from the gastrointestinal tract, nasal passagesand skin surface of the compositions of the invention. Such formulationshave been discovered, and are described below.

SUMMARY OF THE INVENTION

The invention comprises compositions for human use containing one ormore of a flavonoid compound, a non-bovine heavily sulfatedproteoglycan, an unrefined olive kernel extract, a sulfated hexosamine,S-adenosylmethionine (“SAM”), histamine-1 receptor antagonists,histamine-3 receptor agonists, antagonists of the actions of CRH, folicacid, a straight chain polyunsaturated fatty acid, a phospholipid, apolyamine, an interferon and glutiramer acetate, together withappropriate excipients and carriers, said compositions having improvedabsorption from the gastrointestinal tract, skin surface, and nasal andpulmonary surfaces, and anti-inflammatory effects synergistic with eachother and synergistic with available conventional clinical treatmentmodalities.

In one embodiment, the sulfated glucosamine is D-glucosamine sulfate,the proteoglycan is non-bovine chondroitin sulfate, and the flavonoid isquercetin (3,3′,4′,5,7-pentahydroxy flavone), the quercetin glycosiderutin, myricetin, genistein, kaempferol, luteolin, apigenin,(−)-epigallocatechin-3 gallate, kaempferol or the kaempferol glycosideastragaline, or hesperitin or its glycoside hesperidin.

In another embodiment, compositions may also contain antagonists of theeffects of CRH on mast cells or other target cells of the myocardium,gastric mucosa, urinary bladder, skin, meningeal membranes, blood-brainbarrier, and brain structures.

In still another embodiment, the inventive compositions are used againstsuperficial vasodilator flush syndromes.

In still another embodiment, the inventive compositions may be used ascoatings on medical devices, not only to protect surrounding tissuesfrom inflammation due to the devices, but also to treat innateinflammation in surrounding tissues.

In another embodiment, the inventive compositions are used against theinflammatory processes of endometriosis.

In yet another embodiment, the inventive compositions are used againstthe inflammatory components of hormonally-related cancers, such asbreast, ovarian, uterine, prostate and testicular cancers, and whensupplemented with chemotherapeutic agents are used against the canceritself.

In still another embodiment, the inventive compositions may be used inthe treatment of the neuroinflammatory aspects of multiple sclerosis.

In another embodiment, the inventive olive kernel extract is used toimprove the absorption of biochemicals across membrane barriers in thebody, such as those of the intestine, skin, oral mucosa, blood-brainbarrier, and pulmonary alveoli.

In yet another embodiment, the inventive compositions may be used in thetreatment of fibromyalgia or chronic fatigue syndrome.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION

It has been discovered that various combinations of a sulfatedproteoglycan, unrefined olive kernel extract, a flavone (a.k.a.flavonoid compound), a sulfated D-hexoseamine, a phospholipid, a longchain unsaturated fatty acid, a CRH antagonist, a histamine-1 receptorantagonist, a histamine-3 receptor agonist, glutiramer acetate, aninterferon, and a polyamine have synergistic anti-inflammatory effectswhen used as a dietary supplement, a topical product or an aerosol fornasal or pulmonary administration, without or with a conventionalclinical treatment for inflammatory diseases. Within the presentcontext, such inflammatory diseases result from the activation,degranulation and consequent secretion of inflammatory biochemicals frommast cells, and the resultant inflammatory diseases include the groupconsisting of: allergic inflammation, arthritis (to includeosteoarthritis and rheumatoid arthritis), fibromyalgia, chronic fatiguesyndrome, inflammatory bowel disease, interstitial cystitis, irritablebowel syndrome, migraines, atherosclerosis, coronary inflammation,ischemia, chronic prostatitis, eczema, multiple sclerosis, psoriasis,sun burn, periodontal disease of the gums, superficial vasodilator flushsyndromes, hormonally-dependent cancers, and endometriosis. The olivekernel extract alone may be used to improve the transmembrane transportof difficulty-absorbable biomolecules in the intestine, skin andpulmonary alveoli.

In a highly preferred embodiment, the sulfated proteoglycan isnon-bovine chondroitin sulfate, preferably from shark cartilage, whichblocks mast cell activation, degranulation and consequent secretion ofinflammatory biochemicals from the mast cells. Other natural sulfatedproteoglycans suitable for practicing this invention include keratansulfate, dermatan sulfate and hyaluronic acid sodium salt (sodiumhyaluronate). The preferred biological source of the chondroitin sulfateis shark cartilage which is more-highly sulfated than the commoncommercial chondroitin sulfate isolated from cow trachea; the sharkcartilage source also avoids the potential dangers associated withbovine sources.

A highly preferred flavonoid is quercetin which inhibits secretion ofinflammatory molecules from mast cells by affecting moesin, a unique 78kDa mast cell protein [Theoharides, T C, et al., J Pharm. Exp. Therap.294:810 (2000), Kempuraj, et al., Br. J. Pharmacol. 145:934 (2005)]. Inaddition to quercetin, other flavonoids suitable in carrying out theinvention include: the quercetin glycoside rutin, myricetin, genistein,luteolin, apigenin, (−)-epigallocatechin-3 gallate, kaempferol and thekaempferol glycoside astragaline, hesperitin and its glycosidehesperidin.

The olive kernel extract product component of the inventive compositionsis preferably an unrefined (first pressing, filtered, oleic acid-relatedacidity <3%, water content <1%) extract product produced, for onesource, on the island of Crete in Greece. This kernel extract product isespecially prepared by applicant's process consisting essentially of:

(1) harvesting first collection ripe olives, preferably in December; (2)compressing the oil from the flesh of the ripe olives; (3) washing thekernels remaining after step (2) with water to remove debris; (4) dryingthe washed kernels with a stream of hot air; (5) crushing the driedkernels to produce an extract; (6) extracting the extract from step (5)with an organic solvent (e.g., hexane, heptane, octane) plus steam; (7)removing particulate matter from the organic extract by centrifugationor microfiltering through 1-2 micron pore size filters; (8) evaporatingthe organic solvent and water from the clarified extract of step (7) bymaintaining the extract at 86-100 degrees C. while percolating helium(to avoid oxidation) through the fluid, which process reduces the watercontent to <1%, the acidity (as oleic acid) to <3%; and, the organicsolvent to <1%; and (8) storing the final kernel extract product in theabsence of air.

The inventive olive kernel extract surprisingly has the unique propertyof increasing absorption of the other components of theanti-inflammatory compositions through the intestinal mucosa or skin,and also adds its own content of important anti-oxidants, such as omegafatty acids (e.g., eicosapentanoic acid) and alpha tocopherol. Thepolyphenols found in such olive kernel extracts also haveanti-inflammatory effects in, for example, arthritis[Martinez-Dominguez, et al., Inflamm. Res. 50:102 (2001)]. E.B.E.K.,Inc., Commercial, Industrial Enterprises of Crete, 118 EthnikisAntistasecos, Heraklion, Crete, 71306, Greece, or MINERVA SA. Edible OilEnterprises, 31 Valaoriton S1., Metamorphosis, Attizes, Greece willprepare the extract product according to applicant's above-describedprocedure for commercial users.

In addition to its usefulness in increasing the absorption of theinventive macromolecular compositions across the intestinal wall and theskin, the inventive olive kernel extract product is useful in aiding thedissolution of other drugs prior to administration to a patient, and isuseful in promoting the absorption of other difficultly-absorbabledrugs, e.g., the HDL-increasing drug torcetrapib across intestinalmucosa, oral mucosa, nasal mucosa, and skin of patients.

Supplementation of the compositions described above with the methylationreagent S-adenosylmethionine (“SAM”) adds antioxidant, anti-inflammatoryand cytoprotective properties, particularly in inflammatory joint andcardiovascular diseases. Addition of SAM also accelerates metabolism ofhomocysteine, which amino acid has been implicated in coronary disease,to cysteine, which is harmless. Folic acid may be added to certain ofthe present formulations for similar reasons.

Another supplement to the basic compositions of the invention is ahistamine-1 receptor antagonist, such as hydroxyzine, merelastine,azelastine, azatadine, rupatadine, and cyproheptadine. Other histamine-1receptor antagonists are described in Table 25-1 in Goodman and Gilman'sThe Pharmaceutical Basis of Therapeutics, 9^(th) ed., New York, 1996.Histamine-3 receptor agonists are described in the Theoharides patentslisted above.

Inhibitors of mast cell activation and secretion of inflammatorybiochemicals may be used in the treatment of inflammatory processes suchas superficial vasodilator syndrome, such as occurs inmenopausal-associated flush, carcinoid flush, MSG-associated flush, andniacin-associated flush.

Hormone-dependent cancers, including the estrogen/progestin linkedovarian, uterine, breast, and endometrial cancers, and theandrogen-linked prostate and testicular cancers, are associated withtissue inflammation. These conditions can be treated with chondroitinsulfate, quercetin, genistein, rutin, phenoxodiol isoflavone,(−)-epigallocatechin-3-gallate, olive kernel extract, and, optionally,chemotherapeutic agents such as tomoxifen or raloxifen.

Pelvic inflammatory conditions, such as present in endometriosis, canalso be treated with the inventive compositions. Particularly useful inthis regard are compositions delivering 50-300 mg/day of rutin,quercetin, kaempferol, myricetin, or hesperitin.

The inventive compositions may also be used as coatings on implantedmedical devices, which devices may lead to or be associated withinflammation of surrounding tissues, in order to provide protectionagainst such inflammations. Not only can the coating of such medicaldevices inhibit or protect against inflammation caused by the deviceitself, but the coated devices can also be used to deliver the inventivecompositions to innately inflamed tissues due to other causes. Suchmedical devices include artificial skins (scaffolding such as naturallyoccurring polymers, e.g., collagen; man-made polymers, e.g., PTFE,Dacron, PET or polyethylene; self-degrading man-made polymers, e.g., PLAor PGA; biopolymer matrices from animal tissues including fetal andneonatal tissues to be used as tissue engineering scaffolds (cf. Bell etal., U.S. patent application Pub. No. 20020146393)), artificial joints,band-aids, stents for blood vessels, artificial blood vessels,pacemakers, stents for abdominal support in hernia repair, tissuetransplants, prostheses, breast implants, etc. Particularly useful inthis regard are compositions containing heavily sulfated, non-bovineproteoglycans (e.g., chondroitin sulfate) and a flavonoid.

Oral flavonoids, such as those listed above, are reported to influencethe course of experimental autoimmune encephalomyelitis in mice, a modelfor multiple sclerosis (Verbeck, R., et al., Biochem. Pharm. 70(2):220(Jul. 15, 2005); Hendriks, J. J., et al., J Exp Med. 200(12):1667 (Dec.20, 2004). In preferred embodiments of the inventive compositions,flavonoids or flavonoid glycosides plus one or more of a proteoglycan,olive kernel extract, Krill oil, hydroxyzine,(−)epigallocatechin-3-gallate, and a long chain fatty acid plusinjections of interferon and/or glutiramer acetate (Copolymer I)(Copaxone, TEVA Pharmaceuticals, Israel; Avonex, Biogen., USA) are usedin treatment of the chronic inflammation of the central nervous systemin multiple sclerosis. The glutiramer acetate is of particular value inpreventing relapsing/remitting forms of multiple sclerosis [Mezzapesa,D. M., et al., Exper. Rev. Neurother. 5:451 (2005); Schwartz, M., etal., J. Neurol. Sci. 233:163 (2005); Amon, R., et al., Proc. Nat. Acad.Sci. USA 102 Suppl. 2:14593 (2004)].

Sources of CRH antagonists include, in addition to the Theoharidespatents listed in the Background section above: Neurocrine Biochem.Inc.'s D-Phe 12 NIe Ala32,21,38hCRH(12-41)NR2, cat no. 1P-36-41; Pfizernon-peptide CP-154, 526-1; Sigma Chem., St. Louis anti-CRH polyclonalantiserum; and Pfizer, NY patents and applications: U.S. Pat. No.6,211,195, U.S. Pat. No. 5,795,905, PCT/IB95/00573, PCT/IB95/00439, U.S.Ser. No. 08/448,539, U.S. Ser. No. 08/481,413, U.S. Ser. No. 09/735,841,and in Owens et al. Pharm. Rev. 43:425 (1991).

The preferred concentration range of the proteoglycan, hexosaminesulfate, flavonoid, polyunsaturated fatty acid, phospholipid componentsof the oral formulations are 10-3,000 mg per tablet or capsule. Thepreferred concentration range for SAM is 3-1,000 mg per capsule ortablet. Generally, where present, the amounts of the unrefined olivekernel extract are at least twice those of the other active ingredients,preferably 300-1200 mg. The number of capsules or tablets to be takenper day is determined by the nature and severity of the medicalcondition, and is readily determinable by the patient's health provider.Other representative formulations are described in the examples below.

The compositions of the invention may be formulated in any standardmeans of introducing pharmaceuticals into a patient, e.g., by means oftablets or capsules. The compositions of the invention include ointmentsand creams for skin conditions, mouth washes and toothpaste forperiodontal diseases, and solutions for nasal aerosols. Standardexcipients and carriers for the active ingredients of the inventivecompositions are described in Remington's Pharmaceutical Sciences, MackPublishing Co., Easton, Pa.

Although not bound by any particular mechanism of action of thecomponents of the claimed compositions, the inventor contemplates thatthey inhibit the activation and degranulation of the relevant mastcells, and inhibit the secretion of inflammatory biomolecules from thesemast cells. “Activation” and “degranulation” of mast cells are definedherein as is standard and well known in this art, that is, to meansynthesis and secretion from the activated mast cell of any type ofmolecule(s) that alone or in combination triggers inflammation.

EXAMPLES Example 1

Table 1 compares chondroitin sulfate-containing commercial products tothe present compositions.

TABLE 1 Comparison of Chondroitin Sulfate-Containing Products to PresentInvention Product Most Available Compositions Present Invention Mainingredient Mixture of chondroitins Non-bovine chondroitin sulfate,preferably the C type Source Cow trachea Shark cartilage Amount percapsule or tablet 100-300 10-3000 mg Degree of sulfation Low, if anyHigh Absorption from g.i. tract <5% >15% Target Unknown Mast cells,inflammatory cells Other ingredients Vitamins, fish oils (some Flavones,unrefined olive preparations) kernel extract, SAM, histamine-1 receptorantagonists, histamine-3 receptor agonists, CRH antagonists, polyamines,caffeine, folic acid Advantages None known Anti-allergic, anti-inflammatory, antioxidant, cytoprotective Adverse Effects Risk of madcow disease, None known spongiform encephalopathy, stomach upset,allergy to fish products Relevant conditions Osteoarthritis Allergicinflammation angina, asthma coronary artery disease, arthritis(osteoarthritis or rheumatoid arthritis), chronic prostatitis, eczema,fibromyalgia, interstitial cystitis, irritable bowel syndrome,inflammatory bowel disease, migraines, multiple sclerosis, psoriasis,periodontal disease, flush syndrome, cancer (includinghormonally-dependent forms). Scientific publications None foundTheoharides, et al. Br. J. Pharm. 131: 1039 (2000) Middleton, et al.Pharm. Rev. 52: 673 (2000)

In all examples, chondroitin sulfate is assumed to be of a non-bovinevariety.

Example 2

Composition For Protecting Against Inflammatory Diseases Two capsules tobe taken orally 2-3 times daily, at least one hour before mealsIngredients, per capsule, mg: Chondroitin sulfate 150-300 D-Glucosaminesulfate 150-300 Quercetin 150-300 Olive kernel extract  350-1200

Example 3

Composition For Protecting Against Arthritis Ingredients, per capsule,mg: D-Glucosamine sulfate 150-300 Chondroitin sulfate 150-300 Sodiumhyaluronate 100-200 Quercetin 150-300 Olive kernel extract  350-1200

Example 4

Topical Composition For Protecting Against Arthritis Skin ointment orcream. Apply three times per day to affected areas. Ingredients % byweight D-glucosamine sulfate 5 Chondroitin sulfate 5 Sodium hyaluronate0.5 Bitter willow bark extract 5 Quercetin 3 Aloe vera 10 Olive kernelextract 5

Example 5

Composition For Protecting Against Cardiovascular Inflammatory Diseasemg/capsule: *Chondroitin sulfate 50 *Kaempferol 100*S-adenosylmethionine 50 *Niacin 0.01 *Olive kernel extract 350-1200*Bitter willow bark extract 5% by weight *Polyunsaturated fatty 100-600acids (DHA, DPA)

Example 6

Composition For Protecting Against Periodontal Inflammatory DiseaseMouthwash: *Chondroitin sulfate 0.4 M *Quercetin 0.4 M *In a standardmouthwash vehicle

Example 7

Composition For Protecting Against Periodontal Inflammatory DiseaseToothpaste Composition Toothpaste, mg %: *Chondroitin sulfate 5*Quercetin 3 *D-glucosamine sulfate 5 *Olive kernel extract 1 *In astandard toothpaste vehicle

Example 8

Composition For Protecting Against the Inflammation of SunburnIngredients % by weight *Chondroitin sulfate 5 *D-glucosamine sulfate 5*Quercetin 3 *Aloe vera 10 *Olive kernel extract 5 *Sun screen (e.g.,TiO₂) 5

Example 9

Oral Composition For Protecting Against Migraine Headaches Ingredientsmg: *Chondroitin sulfate 50 *Quercetin 100 *Azatadine 4 *Optionally, aCRH-receptor antagonist 5-300

Example 10A

Oral Composition For Protecting Against Inflammation in RelapsingMultiple Sclerosis Ingredients, mg/day *Quercetin 50-300 *Chondroitinsulfate 50-300 *Rutin 50-300 *Hydroxyzine 50-300 *Olive kernel extract350-1200 *Optionally, interferon-beta 8 million IU Betaferon (Schering),s.c., on alternate days or 30 μg Avonex *Optionally, glatiramer acetateCopaxone NPR by parenteral injection

Example 10B

General Composition for Protecting Against the Brain Inflammation ofMultiple Sclerosis Components Mg/tablet or capsule Quercetin 100-1000Rutin 100-1000 (−)Epigallocatechin-gallate 100-1000 Docosohexanoic acid(DHA) 100-1000 Krill oil 100-1000 Olive kernel extract 100-1000

Example 10C

Specific Composition for Protecting Against Multiple SclerosisComponents Amounts Quercetin, rutin, (−)epigallocatechin3-gallate, Each150 mg/tab or cap. Docosohexanoic acid Krill oil  50 mg Olive kernelextract 450 mg

Example 11

Composition For Protecting Against the Inflammation of Cystitis AndProstatitis Ingredients, mg/capsule or tablet: *D-Glucosamine sulfate 50 *Chondroitin sulfate 100-300 *Sodium hyaluronate 200 *Quercetin100-400 *Olive kernel extract  350-1200

Example 12

Composition For Protecting Against Inflammatory “Flush” Ingredients, percapsule: *Chondroitin sulfate 50 mg *Quercetin 150-350 mg *Olive kernelextract 100-750 mg *Bitter willow bark extract 5% by weight *Optionally,cyproheptadine or azatadine 4 mg

Example 13

Cream Composition For Protecting Against Inflammatory Skin AllergyIngredients: % by weight *Aloe vera 5 *Non-bovine chondroitin sulfate 5*Myricetin 5 *Alpha-tocopherol 5 *Olive kernel extract 5 *Aloe vera 10*Optionally, azelastine or hydroxyzine 5

Example 14

Composition For Protecting Against Inflammatory Allergies and AllergicAsthma Ingredients, mg/tablet *Myricetin 500 *Chondroitin sulfate 200*Optionally, azelastine 4 *Rutin 500 *Optionally, hydroxyzine 25

Example 15

Composition For Protecting Against Brain Metastases from Breast CancersIngredients, mg/day Chondroitin sulfate 50-300 Quercetin 25-250Genestein 50-300 Phenoxodiol isoflavone 500-1000 Olive kernel extract350-1200 Optionally, tomoxifen or raloxifen About 10

Example 16

Composition For Protecting Against the Inflammation of AllergicConjunctivitis Ingredients: Weight % *Quercetin 0.05% *Chondroitinsulfate  2.0% *Optionally, azelastine 0.05%

Example 17 Effect of Olive Kernel Extract on Absorption of aProteoglycan Sulfate In Vivo

Chondroitin sulfate was tritiated by New England Nuclear Corp. to aspecific activity of 4.3 mCi/ml.

Unlabeled chondroitin sulfate was dissolved in olive kernel extract at aratio of about 55 w/v chondroitin sulfate powder to about 450 w/v ofolive kernel extract (2.9% acidity as oleic acid, 1.03% water, 0.08%hexane). To this solution was added 20.2 microcuries of the labeledchondroitin sulfate. AAA gelatin capsules were filled with the resultingsolution using an aluminum template molding device.

The laboratory animals (250 g male Sprague-Dawley rats) were keptovernight without food but with free access to water. One capsulecontaining the above-described chondroitin sulfate-olive kernel extractsolution was given to each rat per os. Control animals were given theequivalent amount of chondroitin, but without olive kernel extract. Theanimals were then given free access to food. Serum radioactivity wasmeasured 8 hours thereafter in a beta scintillation counter.

The results showed that, in control animals, about 3.9%+/−0.4% (n=3) ofthe dose of labeled chondroitin sulfate reached the circulation. Insharp contrast, in animals given the olive kernel extract along with thelabeled chondroitin sulfate, about 14.3%+/−0.7% (n=4) of the dose wasabsorbed into the general circulation.

These results demonstrate that olive kernel extract increased by almost400% the absorption of a proteoglycan from the intestine into thegeneral circulation.

Parallel experiments with codfish oil, corn oil and olive oil (from theflesh of the olive) were contemplated, but chondroitin sulfatesolubility in these oils was insufficient to meet the requirements ofthe experiment.

Example 18

Composition for Protecting Against Inflammatory EndometriosisIngredients mg/tablet *Rutin 300 *Quercetin 300 *Olive kernel extract300

1. A method of treating neurodegenerative brain inflammation in asubject, comprising administering to a subject in need thereof aneffective amount of a composition comprising a flavonoid or a flavonoidglycoside, olive kernel oil, a proteoglycan and one or more ingredientsselected from the group consisting of a polyunsaturated fatty acid and aphospholipid.
 2. The method of claim 1, wherein the composition furthercomprises an interferon.
 3. The method of claim 1, wherein thecomposition further comprises glutiramer acetate.
 4. The method of claim1, wherein the composition further comprises a histamine-1 receptorantagonist.
 5. The method of claim 1, wherein the flavonoid is selectedfrom the group consisting of quercetin, myricetin, genistein,kaempferol, (−)epigallocatechin-3-gallate, luteolin, epigenin, rutin,hesperitin, and hesperidin.
 6. The method of claim 1, wherein thecomposition comprises a polyunsaturated fatty acid, and wherein thepolyunsaturated fatty acid is selected from the group consisting ofalphalinoleic acid, eicosapentenoic acid, and docosahexenoic acid. 7.The method of claim 1, wherein the composition comprises a phospholipid,and wherein the phospholipid is selected from the group consisting offish oil, Krill oil, and sphingomyelin.
 8. The method of claim 1,wherein the composition comprises chondroitin sulfate, a flavonoid,docosahexenoic acid, Krill oil, and olive kernel oil.
 9. The method ofclaim 8, wherein the composition further comprises hydroxyzine.
 10. Themethod of claim 8, wherein the composition is administered orally. 11.The method of claim 1, wherein each ingredient is in the amount of100-1,000 mg.
 12. The method of claim 1, wherein the compositioncomprises chondroitin sulfate, quercetin, rutin,(−)epigallocatechin-3-gallate, docosahexenoic acid, and olive kerneloil.
 13. The method of claim 12, wherein the composition comprises 150mg of quercetin, 150 mg of rutin, 150 mg of(−)epigallocatechin-3-gallate, 250 mg of docosahexenoic acid, 300 mg ofolive kernel oil and chondroitin sulfate.
 14. The method of claim 12,wherein the composition further comprises hydroxyzine.
 15. The method ofclaim 12, wherein the composition is administered orally.
 16. The methodof claim 1, wherein the neurodegenerative brain inflammation includesdisruption of the blood-brain barrier.
 17. A composition comprising10-1000 mg of luteolin, 10-1000 mg of quercetin, 10-1000 mg of rutin,100-1000 mg of (−)epigallocatechin-3-gallate, 100-1000 mg ofdocosahexenoic acid, 100-1000 mg of krill oil, and 100-1000 mg of olivekernel oil.
 18. The composition of claim 17, wherein the compositionfurther comprises 10-1000 mg of chondroitin sulfate.
 19. The compositionof claim 18, wherein the composition comprises 150 mg of luteloin, 150mg of quercetin, 150 mg of rutin, 150 mg of(−)epigallocatechin-3-gallate, 250 mg of docosahexenoic acid, 300 mg ofolive kernel oil and 10-1000 mg of chondroitin sulfate.